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ER Reorganization is Remarkably Induced in COS-7 Cells Accumulating Transmembrane Protein Receptors Not Competent for Export from the Endoplasmic Reticulum

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Abstract

The newly synthesized mutant L501fsX533 Frizzled-4 form and the alpha3beta4 nicotinic acetylcholine receptor expressed in the absence of nicotine accumulate in the endoplasmic reticulum of COS-7 cells and induce the formation of large areas of smooth and highly convoluted cisternae. This results in a generalized block of the transport to the Golgi complex of newly synthesized proteins. Intriguingly, both effects happen peculiarly in COS-7 cells; HeLa, Huh-7, and HEK293 cells expressing the two receptors at similar level than COS-7 cells show normal ER and normal transport toward the plasma membrane. These results question the conclusion that a dominant-negative mechanism would explain the dominance of the mutant L501fsX533 Fz4 allele in the transmission of a form of Familial exudative vitreoretinopathy. Moreover, they indicate that the coordination of endoplasmic reticulum homeostasis in COS-7 cells is particularly error prone. This finding suggests that COS-7 cells may be extremely useful to study the molecular mechanisms regulating endoplasmic reticulum size and architecture.

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Acknowledgments

This work was supported in part by the Telethon (Grant GGP09029 to SB) and the CNR Research Project on Aging, Regione Lombardia Projects NUTEC ID 30263049 and MbMM-convenzione no. 18099/RCC (SFC and AC). We thank the Monzino Foundation (Milan, Italy) for its generous gift of the Spinning Disk confocal system LCI MLS (PerkinElmer). We thank Cecilia Gotti for the alpha3 and beta4 nicotinic subunit antibodies, Gabriella Caporaso for her continuous support throughout the Project and Nica Borgese for suggestions.

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Correspondence to Sara Francesca Colombo or Stefano Bonatti.

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D’Agostino, M., Crespi, A., Polishchuk, E. et al. ER Reorganization is Remarkably Induced in COS-7 Cells Accumulating Transmembrane Protein Receptors Not Competent for Export from the Endoplasmic Reticulum. J Membrane Biol 247, 1149–1159 (2014). https://doi.org/10.1007/s00232-014-9710-8

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